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Genes in Bottle Kit
Genes in Bottle Kit
Catalog Number
166-2000EDU
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Capture Your Essence!
Bottle your DNA! Whether it’s being cloned, sequenced, fingerprinted, mapped, or
genetically engineered, DNA has become an everyday topic in the media and the classroom.
Introduce your students to the molecular framework of biology — with their own DNA!
How do scientists separate pure DNA from cells composed mainly of lipids, proteins,
carbohydrates, and salts? Membranes are first ruptured with detergents to release DNA
into a solution; then proteins and other organic molecules are digested and separated while
retaining intact DNA. The DNA is finally collected by precipitation in a form that can be
manipulated as desired.
With this simple lab activity, students gain practical knowledge by conducting a real-world
procedure that is used to extract DNA from many different organisms for a variety of
applications. Your students will extract genomic DNA from their own cheek cells and watch
it precipitate from solution as floating white strands. The DNA strands are then easily
collected and transferred to a glass vial, and the vial is fashioned into a necklace!
Seeing is believing. For students learning about the molecular framework of biology for
the first time, DNA is abstract and intangible. This procedure makes the invisible visible —
seeing their own DNA makes it real and helps students comprehend this previously invisible
substance of life.
Learning opportunities for all levels of instruction. This activity is designed for any
classroom environment and requires no specialized equipment or stains. For secondary
and college level instruction, lessons on DNA structure and function, cell structure, and
enzyme function can be introduced or reinforced with this laboratory activity. For middle
school students, it’s a perfect introduction to the exciting world of DNA science.
Environmental
Scientific
and Health
Inquiry
Science
• Genetic testing
• DNA fingerprinting
Genes in
Chemistry
a Bottle of Life
Kit • Chemical properties of
cell components
• Cell structures • Properties of enzymes
Structure Heredity • Solubility
• Organelles
• Nuclear and DNA and Function and Molecular
staining of Organisms Biology
• Cell organization
Student Manuals
Basic Level Student Manual ........................................................................................13
Introduction..............................................................................................................14
Workstation Checklist ..............................................................................................17
Procedure for DNA Extraction and Precipitation ......................................................17
Advanced Level Student Manual ................................................................................21
Introduction and Focus Questions ..........................................................................22
Workstation Checklist ..............................................................................................27
Procedure for DNA Extraction and Precipitation ......................................................27
Extension Activities
Dry Laboratory Demonstration of DNA Extraction ....................................................30
Microscropic Observation and Nuclear Staining of Cheek Cells ..............................30
Staining precipitated DNA........................................................................................31
Answers to Focus Questions (Basic Instruction) ........................................................32
Answers to Focus Questions (Advanced Instruction)..................................................33
Teacher’s Guide
Kit Inventory Checklist
This section lists the components provided in this Genes in a Bottle Kit. It also lists required
and optional accessories. Each kit contains sufficient materials to outfit 9 student workstations
of up to four students per workstation. Use this checklist to inventory your supplies before
beginning advanced preparation.
Kit Components Quantity ✔)
(✔
Lysis buffer 150 ml ❐
Powdered protease and salt 1.5 g ❐
15 ml conical tubes 50 ❐
Clear micro test tubes 60 ❐
Multicolor micro test tubes 60 ❐
Disposable plastic transfer pipets 60 ❐
Foam micro test tube holders 10 ❐
Required Accessories (not included in this kit) Quantity ✔)
(✔
91% isopropanol (available at drug stores) approx. 360 ml ❐
or 95% ethanol
Water bath with thermometer, set at 50°C* 1 ❐
Permanent markers 1–9 ❐
Container of ice 1 ❐
Disposable paper cup or beaker for waste disposal 9 ❐
Beaker or rack to hold 15 m tubes in water bath
(need space for 36 tubes maximum) 1 ❐
Optional DNA Necklace Module** (not included in this kit)
**Each DNA necklace module contains enough material to prepare 18 necklaces. Two kits
are required for a class of 36 students. 166-2200EDU contains:
Glass vials 18
Silver caps 18
Plastic plugs 18
Waxed string 18
Super glue gel 1 tube
* If a temperature-controlled water bath is not available, use one or more insulated
containers (Styrofoam is best) large enough to hold a beaker or rack containing up to 36
15 ml tubes, and fill with water heated to 50°C.
Catalog # Description
166-2300EDU Genes in a Bottle Kit, contains (1) DNA Extraction Module and (2)
DNA Necklace Modules. Serves up to 36 students
166-2000EDU Genes in Bottle DNA Extraction Module (serves 36 students)
166-2200EDU Genes in a Bottle DNA Necklace Module (serves 18 students)
166-2001EDU Genes in a Bottle DNA Extraction Refill Package, includes lysis
buffer and powdered protease + salt
166-2002EDU Genes in a Bottle Lysis Buffer, 150 ml
1
Cheek Cell DNA Extraction
Intended Audience
This laboratory is appropriate for students from 5th grade through college, as a first
introduction to DNA or as a quick, easy, and impressive hands-on accompaniment to
existing DNA instruction. Even students who have previously extracted DNA out of onions
or liver will find extracting their own DNA far more relevant and exciting.
The instruction manual includes content for both advanced instruction (9th grade through
college) and basic instruction (5th through 8th grades). Depending on the needs of your
students, you may choose to include activities or background material from either section.
A complete student manual is provided for both levels of instruction.
2
Curriculum Fit
This laboratory activity can be performed at any point during a typical biology or life science
year, but it is particularly relevant when the following topics are being discussed:
• Biomolecules
• Cell structure
• Mitosis and meiosis
• Genetics
• DNA technology
Activity Timeline
This laboratory activity can be performed easily in one 45-minute class period but can be
expanded to include several extension activities.
Lesson 1 Introduction and background material
Lesson 2 Cheek cell isolation, DNA extraction, and precipitation
Lesson 3 DNA necklace preparation (optional)
Safety Issues
In this experiment, no special biosafety handling is required. There is no greater risk of
exposure to infectious agents in this activity than in normal student interactions (sharing a
beverage, sneezing). Students will handle their own biological samples. Lysis buffer is
added to break open the cells, rendering them inviable.
Eating, drinking, smoking, and applying cosmetics are not permitted in the work area.
Wearing protective eyewear and gloves is strongly recommended. Students should wash
their hands with soap before and after this exercise. If any of the solutions gets into a student’s
eyes, flush with water for 15 minutes.
Keys to Success
Ample cell collection is critical for success. For best results, make sure students spend the
recommended amount of time collecting and carefully transferring cheek cells.
Volume Measurements
This kit was developed for use in classrooms with minimal laboratory equipment and limited
knowledge of scientific techniques. Micropipets are not required but can be used to transfer
liquids.
3
Background and Fundamentals for Basic Level
Instruction
What is DNA and what does it do?
Deoxyribonucleic acid (DNA) is a molecule present in all living things, including bacteria,
plants, and animals. DNA carries genetic information that is inherited, or passed down from
parents to offspring. It is sometimes referred to as a biological “blueprint“ because it
determines all of an individual’s physical features such as hair, eye, and skin color, height,
shape of facial features, blood type, and countless others. Your DNA blueprint is a
combination of your mother’s DNA (from her egg) and your father’s DNA (from his sperm)
during conception.
DNA contains four chemical units, referred to by the first letters in their names: A (adenine),
G (guanine), T (thymine), and C (cytosine). These four letters make up a code for genetic
information. The letters of the DNA code function like letters of our alphabet. The 26 letters
in the English alphabet spell words, which can be arranged in infinite ways to create
messages and information. Similarly, the 4 chemical letters of DNA are organized to make
messages that can be understood by cells, called genes. These genes contain the
information to make proteins, which are the basis for almost all of a body’s and cell’s
structures and functions.
Your DNA sequence is the particular arrangement or order of the chemical letters within
your complete DNA collection, or genome. Scientists have determined that human DNA
sequences are 99.9% identical. It is the <0.1% sequence variation from person to person
that makes each of us unique.
4
What does DNA look like?
At the molecular level, DNA looks like a twisted ladder or a spiral staircase. The ladder
actually contains two strands of DNA, with pairs of the chemical letters A, G, T, and C
forming the rungs. This structure is called a DNA double helix because of the spiral, or
helical form made by the two DNA strands. Each strand of DNA is very long and thin and is
coiled very tightly to make it fit into the cell’s nucleus. If all 46 chromosomes from a human
cell were uncoiled and placed end to end, the DNA would be 2 meters long — but only 2
nanometers (2 billionths of a meter) wide.
5
Background and Fundamentals for Advanced Level
Instruction
Applications of DNA Technology
This laboratory activity can be integrated into classes that discuss DNA structure and function
and can be used to give students a simple, hands-on experience with their own DNA. It
takes on even more significance if students understand that DNA extraction is the first step
of many biotechnology applications, such as:
Cloning
Cloning means to make many copies of a fragment of DNA or genome. A defective gene
that causes disease may be cloned so that it can be sequenced and analyzed toward the
goal of finding a cure. A gene encoding a desirable protein or trait may be cloned so that it
can be inserted into another organism (see Gene Transfer below). Likewise, an entire
genome can be cloned by inserting it into cell nuclei that are capable of developing into
organisms.
DNA Profiling
Using a technique called the polymerase chain reaction (PCR), scientists can study specific
regions of chromosomes where individuals’ DNA sequences differ, and amplify, or make
many copies of them (creating sufficient quantities of these sequences to manipulate and
analyze). Using gel electrophoresis, the differences between individuals can be displayed
as banding patterns that resemble bar codes. This technique can be used to solve crimes,
test paternity, and also to determine the evolutionary relatedness of organisms.
6
DNA and other cellular components, such as fats, sugars, and proteins, dissolve in the lysis
buffer. DNA has a negative electrical charge due to the phosphate groups on the DNA
backbone, and the electrical charge makes it soluble. When salt is added to the sample, the
positively charged sodium ions of the salt are attracted to the negative charges of the DNA,
neutralizing the electrical charge of the DNA. This allows the DNA molecules to come
together instead of repelling each other. The addition of the cold alcohol precipitates the
DNA since it is insoluble in high salt and alcohol. The DNA precipitate starts to form visibly
as fine white strands at the alcohol layer boundary, while the other cellular substances
remain in solution.
7
Teacher’s Laboratory Guide
This section presents an overview and lesson flow, advance preparation,
student workstation setup, and techniques and concepts to highlight.
Implementation Timeline
1–2 days Lesson 1 Introduction and background material
Optional dry laboratory demonstration of DNA
extraction — recommended for students in grades 5–8.
See extension activities at the end of the manual.
45 minutes Lesson 2 Cheek cell isolation, DNA extraction, and precipitation
30–45 minutes Lesson 3 Optional DNA necklace preparation
1 ml
1ml
750 µl
500 µl
250 µl
100 µl
• Place the alcohol (isopropanol or ethanol) in the freezer at least 1 hour before beginning
this laboratory.
• Take the pouch containing the powdered protease + salt (‘prot’) and cut open one
corner. Pour the powder into one of the 15 ml tubes. Add 15 ml of water to the
prot. Drinking water works well; distilled water, as used in laboratories, may be
acceptable.
Once the prot is rehydrated, it is good for up to a week if stored in a refrigerator, at 4°C.
If you plan to use the kit for several groups of students over a few weeks, it is
recommeded that you measure out some of the protease for use now, and rehydrate
the remaining protease for use later. The protease should be rehydrated at a
concentration of 100 mg/ml.
Aliquot 1.25 ml of the rehydrated prot into 8 pink micro test tubes as described below.
8
Aliquotting of Solutions for Each Student Workstation (4 students/station)
1. For each student, dispense 3 ml of water into a 15 ml tube (up to 4 tubes per
station). Any type of drinking water is acceptable.
2. Dispense 1.25 ml of the rehydrated protease + salt (see p. 8 for dilution instructions) into
9 pink test tubes and label the tubes “prot”.
3. Dispense 10 ml of lysis buffer into 9 x 15 ml tubes. Label each tube “lysis”.
4. Place 4 x 15 ml tubes of water and one tube of lysis buffer in a cup or test tube holder,
and 1 pink micro test tube labeled “prot” in a foam micro test tube holder at each student
workstation.
Note: Some users may find collecting mouthwash in 15 ml tubes difficult. As an alternative,
instructors may elect to use a small drinking cup to dispense water and collect mouthwash.
9
DNA Extraction and Precipitation
Workstation Checklist
The materials in this kit are sufficient for 36 students.
10
Quick Guide for DNA Extraction and Precipitation
6. Place the cap on the tube, and gently invert the tube
5 times (don’t shake your tube!). Observe your tube
— do you notice any changes? If you do, write them
down.
11
8. Place the cap on your tube, and gently invert it a few
times.
11. Place your cap on your tube, and let it sit undisturbed
for 5 minutes. Write down anything you observe
happening in the tube.
12
Student Manual: Basic Instruction
Contents
Lesson 1 Introduction and background material, dry laboratory
extension (optional)
Lesson 2 Cheek cell isolation, DNA extraction, and precipitation
Lesson 3 DNA necklace preparation (optional)
13
Student Manual: Basic Instruction
Cheek Cell DNA Extraction
Capture Your Genetic Essence in a Bottle
Introduction
What is DNA and what does it do?
Deoxyribonucleic acid (DNA) is a molecule present in all living things, including bacteria,
plants, and animals. DNA carries genetic information that is inherited, or passed down from
parents to offspring. It is responsible for determining a person’s hair, eye, and skin color,
facial features, complexion, height, blood type, and just about everything else that makes
an individual unique. But it also contains all the information about your body that is the
same in all human beings. In other words, your DNA is like a blueprint for your entire
physical growth and development. Your DNA blueprint is a combination of half of your
mother’s and half of your father’s DNA, which is why you have some features from each of
your parents.
DNA contains four chemical units, referred to by the first letters in their names: A (adenine),
G (guanine), T (thymine), and C (cytosine). These four DNA “letters” make up a code for
genetic information. The letters of the DNA code are similar to the letters of our alphabet.
The 26 letters in our English alphabet spell words, which can be arranged in infinite ways to
create messages and information. Similarly, the 4 chemical letters of DNA are organized to
make messages, called genes, that can be understood by cells. These genes contain the
information to make proteins, which are responsible for almost all of your body’s structures
and functions. A gene is like a recipe, since it contains the all the information needed to
make a protein.
Your DNA sequence is the particular arrangement or order of the chemical letters within
your complete DNA collection, or genome. Scientists have determined that human DNA
sequences are 99.9% identical. It is the <0.1% sequence variation from person to person
that makes each of us unique. In other words, what makes you different from your classmate
is an occasional difference in the letters of your genomes.
Fig. 2. A schematic representation of DNA (deoxyribonucleic acid). DNA is a long chainlike molecule that
stores genetic information.
Workstation Checklist
1 ml
1ml
750 µl
500 µl
250 µl
100 µl
6. Place the cap back on your tube. Gently invert your tube 5 times to lyse your cells.
Don’t shake the tube. If you observe any changes to your cells at this time, write them
down.
2. Place your cell extract tube in the beaker or test tube holder in the 50°C water bath (at
the common workstation) for 10 minutes to allow the protease to work.
Water bath
2. Obtain the tube of cold alcohol from your instructor or at the common workstation. Add
10 ml of the alcohol to your tube as follows. Hold your tube at a 45 angle and add the
alcohol by slowly dispensing it down the inside wall of the tube. It will take repeated
additions to add 10 mls. Screw the cap back onto your tube.
3. Place your 15 ml tube upright either in the cup or a test tube rack and leave it
undisturbed at room temperature for 5 minutes.
4. After 5 minutes, look again at the contents of your tube, especially in the area where
the alcohol and cell extract layers meet. Do you see anything? Write down your
observations. Compare your sample with those of your classmates.
5. With the cap of your tube tightly sealed, mix the contents of your tube by slowly inverting
the tube 5 times. Look for any stringy, white or clear material. This is your DNA!
or
Contents
Lesson 1 Introduction and background material
Lesson 2 Cheek cell isolation, DNA extraction, and precipitation
Lesson 3 DNA necklace preparation (optional)
Introduction
Deoxyribonucleic acid (DNA) is a molecule present in all living things, including bacteria,
plants, and animals, and in almost all cell types. DNA is the carrier of genetic information
and is responsible for determining a person’s hair, skin, and eye color, facial features,
complexion, height, blood type, and just about everything else that makes an individual
unique. It also carries information required for cells to perform all of the functions that are
common to all members of a species, or to all living things, and thus it is sometimes
referred to as a biological “blueprint”. Your personal blueprint is a combination of half of
your mother’s DNA (from her egg) and half of your father’s DNA (from his sperm) during
conception. All of your cells contain this complete set of instructions.
All DNA looks the same when it is extracted from cells, but it is exciting to look at your own
DNA, knowing that this is really what makes you unique and alive. In this laboratory activity,
you will extract your own DNA — a substance that holds your very own “blueprint” — from
your cheek cells. You will use a quick and easy procedure that scientists routinely use to
extract DNA from different organisms.
Every day scientists are making new discoveries as they study the information encoded in
our DNA. Understanding DNA holds the possibility of curing diseases, the hope for millions
who suffer from various genetic disorders and syndromes, making better products from
biological sources, and even perhaps the key to longer life. We are beginning to understand
who we are and why by studying our genetic material.
DNA Structure
At the molecular level, DNA looks like a twisted ladder or a spiral staircase. Two long
molecules are aligned with each other, and the rungs are formed from pairs of chemical
units called bases. This structure is referred to as a double helix because of the spiral, or
helical form made by two strands. The bases function like letters in a code, so they are
known as A, G, T, and C (abbreviations for their full names, adenine, guanine, thymine,
and cytosine, respectively). Each base is
connected to a sugar and a phosphate group, and the sugar and phosphate groups form the
“backbones” of the ladder-like structure. (A nucleotide is one unit consisting of a base,
sugar, and phosphate.) Scientists have found that A always pairs with T, and G always
pairs with C in double-stranded DNA.
The 4 chemical letters of DNA are organized to make messages that can be understood by
cells, called genes. These genes contain the information to make proteins, which are the
basis for almost all of your body’s structures and functions. Each of your cells contains
several billion letters of DNA “text”.
A DNA sequence is the particular arrangement or order of the bases along the DNA
molecule. Human DNA sequences are 99.9% identical among each other. It is the <0.1%
sequence variation that makes each of us unique. In other words, what makes you different
from your classmate is an occasional difference in the sequence of bases in your genes.
3. If you wanted to isolate a copy of the gene that codes for a protein found in the stomach,
could that gene be located in cheek cells? Explain your reasoning.
6. Why is an intermediate like mRNA needed to copy the information from the genomic
DNA so it can be translated into proteins?
7. What do you think will be the first step in isolating DNA from your cells?
Step 2. Lysing the cells and dissolving the phospholipid bilayer membranes
If you guessed that the first step of DNA extraction is to break open the cells, you are right!
Detergent dissolves hydrophobic (oil-based) molecules, and the cell and nuclear membranes
are mainly oil-based (you may have already heard of cell membranes being composed of
“phospholipid bilayers”). After collecting cells from your cheeks, you will add a solution that
contains detergent.
Focus questions:
8. Once the membranes have been dissolved, the DNA is released into the solution, but
so are many other types of cellular molecules. List some types of molecules besides
DNA that you would expect to find in a cell.
9. What method or agent do you think might be used to break down these unwanted
molecules?
11. The protease used in this procedure functions best at 50°C. Would you expect this
enzyme to be isolated from E. coli bacteria? Explain your answer. Hint: Where does
E. coli live?
12. Meat tenderizer is often used to tenderize tough pieces of meat, like steak. Knowing
that steak is made of protein-rich muscle tissue from cows, can you think of an
explanation for how meat tenderizer works?
Student Manual: Advanced Instruction
25
Step 4. Making DNA insoluble
The protease solution that you added to your sample also contains salt. The salt will cause
the DNA to become less soluble in solution. DNA has a negative electrical charge due to the
phosphate groups on the DNA backbone. When the salt is added, the positively charged
sodium ions of the salt are attracted to the negative charges of the DNA, neutralizing the
electrical charge of the DNA. This allows the DNA molecules to come together or
aggregate instead of repelling each other.
Step 5. Precipitating the DNA with cold alcohol
To separate the DNA from the other molecules in the cell extract, you will add cold alcohol
to your sample. Upon the addition of cold alcohol, the DNA will precipitate because it is less
soluble in alcohol than in water. The colder the ethanol is, the less soluble the DNA will be
in it. This is similar to the solubility of sugar in tea (or any drink); sugar dissolves more
readily in hot tea than in iced tea.
In the presence of high salt and cold alcohol, the DNA that had been released from your
cells precipitates and aggregates until it can be seen with the naked eye! The other
molecules in the cell extract, such as the amino acids and carbohydrates, remain dissolved
in the alcohol and water and will not be visible. It takes many thousands of strands of DNA
to form a fiber large enough to be visible. Each strand will have thousands of genes on it,
so you will be looking at material that contains millions of genes at once. Remember,
though, that you are seeing the DNA from many thousands of cells all together.
Focus questions:
13. Match the outcomes on the left with the laboratory steps on the right.
Harvest the cells A. Gently chew the insides of your mouth and
then rinse vigorously with water
Workstation Checklist
1 ml
1ml
750 µl
500 µl
250 µl
100 µl
6. Place the cap back in your tube. Gently invert your tube 5 times to lyse your cells. Don’t
shake the tube. If you observe any changes to your cells at this time, write them down.
2. Place
your cell extract tube in the beaker or test tube holder in the 50°C water bath (at the com-
mon workstation) for 10 minutes to allow the protease to work.
Water bath
28
2. Tilt your 15 ml tube at a 45° angle and slowly add the alcohol, carefully letting it flow
gently down the inside of the tube. Fill the tube with cold alcohol (about 10 ml total).
You may need to use several pipets full of cold alcohol. You should be able to see two
layers (upper and lower) forming. As you add the alcohol, pay close attention to the
place where the alcohol and cell extract layers meet. Write down your observations.
3. Place your 15 ml tube upright either on the cup or a test tube and leave it undisturbed
at room temperature for 5 minutes.
4. After 5 minutes, look again at the contents of your tube, especially in the area where the
alcohol and cell extract layers meet. Do you see anything? Write down your observations.
Compare your sample with those of your classmates.
5. With the cap of your tube tightly sealed, mix the contents of your tube by slowly inverting
the tube 5 times. Look for any stringy, white or clear material. This is your DNA!
6. If you are going to make a DNA necklace, your teacher will provide you with a glass
vial. With a disposable plastic transfer pipet, carefully transfer the precipitated DNA
along with approximately 750 µl to 1 ml of the alcohol solution into the vial. Your teacher
will help you seal the vial so you can complete the necklace.
If you are not going to make a DNA necklace, you can transfer and save your DNA in a
fliptop micro test tube. With a disposable plastic transfer pipet, gently withdraw your
precipitated DNA along with about 1 ml of alcohol solution and transfer it into the micro
test tube. Tighten the cap and amaze your friends and family with your own DNA.
or
Objective
Prepare 1:50 dilution of Fast Blast stain (and 1x PBS, if necessary)*
Required Materials
• 500x Fast Blast DNA stain (catalog #166-0420EDU)
• 200 µl micropipet or disposable plastic pipet tips
• Micropipet tips (if using a micropipet)
• 5 ml isotonic saline solution (e.g., contact lens saline solution or 1x PBS)*
Procedure
Dilute the 500x Fast Blast DNA stain before using it to visualize the nuclei of cheek cells.
To prepare 5 ml of diluted stain, add 100 µl of 500x Fast Blast to 4.9 ml of isotonic saline
solution.
Note: An isotonic solution is necessary to maintain the proper osmotic balance across the
cell membrane.
* 10x PBS can be purchased (#166-2403EDU). Dilute to 1x using distilled water. Alternatively, prepare 1x PBS by dissolving 8 g of
NaCI, 0.2 g of KCI, 1.44 g of Na2HPO4, and 0.24 g of KH2PO4 in 800 ml of distilled water. Adjust the pH to 7.4 with HCl or NaOH
and then bring the volume to 1 L with distilled water. Sterilize by autoclaving or by filtering, and store at room temperature.
30
Required Equipment and Accessories
• Microscope
• Microscope slides
• Glass coverslips
• Disposable plastic transfer pipet or dropper
• Sterile micropipet tips*
• 20 µl micropipet
Procedure
Collect cheek cells
Collect cheek cells by gently scraping the inside of your cheeks 10 times with a sterile pipet
tip.* This is most easily done by pinching and extending the corner of your mouth with one
hand, and scraping the cheek with the tip in the other hand. Use firm but gentle pressure.
You should see a volume of white cells at the end of the pipet tip.
Stain cells
1. Add a drop of diluted Fast Blast to the microscope slide. Place the pipet tip with the
cheek cells on the end of the a micropipet that is set to 20 µl. Gently pipet up and down
5 times to transfer your cheek cells into the drop of Fast Blast stain.
2. Cover the sample with a clean coverslip. View the slide under a microscope at low and
medium power magnifications.
Note: Students should have had prior instruction in the proper use of a microscope. Fast
Blast stains cell nuclei within 2 minutes. Have your students make observations and draw
sketches, labeling visible cell structures.
* If a micropipet and tip are not available, a cytology brush or cotton swab can be used to collect and mix the cells into the diluted
Fast Blast stain.
31
Answers to Focus Questions (Basic Instruction)
1. How could you test whether you were actually collecting cells from your cheeks?
What piece of laboratory equipment might you use?
You could touch your brush to a glass microscope slide after collecting your cheek cells
and look at them under a microscope.
2. When washing dishes, what works better, warm or cold water? Which do you
think will help the detergent break open the cell, warm or cold temperature?
Warm water works better when washing dishes because it helps make the fats and
proteins dissolve better in dish detergent. Warm temperature will help the detergent in
the lysis buffer break open the cells.
3. Do you think your DNA will be visible after you have broken open your cells?
Why or why not?
Your DNA will not be visible after you have broken open your cells. It will be dissolved
in the lysis buffer.
4. Where do you think you would find proteases in your body? Hint: Where do the
proteins that you eat get broken down?
Proteases are found in your stomach, where the proteins that you eat get digested.
5. Have you ever tried to add sugar to iced tea? Does the sugar dissolve easily?
How does this compare to dissolving the same amount of sugar in the same
amount of hot tea?
Sugar dissolves much less easily in iced tea than in hot tea. The cold temperature of the
iced tea reduces the sugar’s solubility, or ability to dissolve. In general, heat increases
the solubility of substances dissolved in liquid.
32
Answers to Focus Questions (Advanced Instruction)
1. Imagine you are trying to explain the difference between chromosomes, genes,
and DNA to your younger brother or sister who is two years younger than you.
Write down your explanation in simple words that they could understand.
DNA is a chemical found in all living things and is passed from parents to children. It
carries the information needed to make you who you are.
Chromosomes are long strands of coiled DNA. The DNA within your cells is organized
into structures called chromosomes, which make it easy to store within the cell and to
copy when cells divide.
Genes are sections of DNA that contain the information needed to make proteins,
which perform critical jobs within living cells.
2. Does a liver cell contain the same chromosomes as a cheek cell?
Yes. The genomic DNA found in all nonreproductive cells is the same, no matter what
tissue the cells come from.
3. If you wanted to isolate a copy of the gene that codes for a protein found in the
stomach, could that gene be located in cheek cells? Explain your reasoning.
The gene that codes for a stomach protein would be found in the genomic DNA inside
a cheek cell. However, the cheek cell would not make the messenger RNA, or copies,
of the gene for the stomach protein. Stomach protein genes are expressed only in the
stomach.
Below is a schematic image of a cheek cell.
Nucleus
Mitochondria
Endoplasmic
reticulum
Plasma (cell)
membrane
Golgi
apparatus Cytoplasm
4. Label the cellular compartments, including the cell membrane, cytoplasm, and
nucleus.
33
6. Why is an intermediate like mRNA needed to copy the information from the
genomic DNA so it can be translated into proteins?
Genomic DNA is in the nucleus and always remains there (like an archived book that
can never leave a library), but the protein-making ribosomes are in the cytoplasm. A
mobile intermediate is needed to bring the genetic information from the nucleus to the
cytoplasm.
7. What do you think will be the first step in isolating DNA from your cells?
The cell and nuclear membranes must be disrupted to release the DNA.
8. Once the membranes have dissolved, the DNA is released into the solution, but
so are many other types of cellular molecules. List some types of molecules
besides DNA that you would expect to find in a cell.
Proteins, lipids, sugars, and minerals (salts) are common cell components.
9. What method or agent do you think might be used to break down these unwanted
molecules?
There are enzymes that specifically digest all kinds of biological molecules. Proteases
break down proteins, detergents dissolve lipids, and enzymes like beta-galactosidase
break down sugars. Heat and agitation can speed up these digestion processes.
10. What proteins might be associated with DNA in the cell?
Chromosomal DNA is bound by histones. Other associated nuclear proteins may
include DNA polymerase or transcription factors.
11. The protease used in this procedure functions best at 50°C. Would you expect
this enzyme to be isolated from E. coli bacteria? Explain your answer. Hint:
Where does E. coli live?
No. E. coli, which lives in our gut, thrives around our body temperature, 37°C. An
enzyme whose optimal temperature is 50°C was probably isolated from an organism
that lives at or near that temperature.
12. Meat tenderizer is often used to tenderize tough pieces of meat, like steak.
Knowing that steak is made of protein-rich muscle tissue from cows, can you
think of an explanation for how meat tenderizer works?
Many meat tenderizers contain papain, which is a protease. The protease breaks down
the protein molecules. By partially degrading some of the proteins, the tough
muscle/meat is made softer and more tender.
13. Match the outcomes on the left with the laboratory steps on the right.
A Harvest the cells A. Gently chew the insides of your mouth and
then rinse vigorously with water
C Dissolve cell membranes B. Add protease, incubate at 50°C
D Precipitate the DNA C. Mix in a detergent solution
B Break down proteins D. Layer cold alcohol over cell extract
E Make DNA less soluble in water E. Add salt
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Bio-Rad
Laboratories, Inc.
4110034 Rev B